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We narrowed to 520 results for: HA tag

Showing: 261 - 280 of 520 results

  1. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ... an essential aspect of biological research that has defined our understanding of biological processes...model organisms is Escherichia coli. The bacterium has been used as a model organism for over 60 years with...cloning and metabolic engineering.  The E. coli genome has an extensive research history and characterization...studies. For example the E. coli biotin ligase, BirA, has been used to identify and study protein-protein interactions...APEX (Rhee et al., 2013). Research using E. coli has the potential to address many areas of microbial ...research with an archive of information and data that has yet to be produced with mice (Iannaccone et al., ..., 2009). While the rat as a model organism has its perks, it is far from perfect. One of the most limiting...

  2. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...technology has been studied and used in assays for over half a century; the Addgene blog has an excellent...new aptamer also has a higher melting temperature than its Spinach ancestors, and has a lower magnesium...initially still unstable in eukaryotes, the Jaffrey lab has since developed new RNA-based scaffolds to contain... As biosensors, aptamers hold a few important advantages over their amino acid-based counterparts, antibodies...acceptor FRET know-how with aptamers, aptabeacons (or aptatags) were born. Aptabeacons can be created in many...quenchers are still relatively bulky. One of the advantages of oligonucleotides over proteins is their modularity... Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the nature of GFP fluorescence...

  3. Twenty Years of Addgene Sharing: CRISPR

    Type
    Blog Post
    ...Repair has been viewed 219,289 times, while Validating Your Genome Edit has 76,011 views.  Has all of ...libraries Addgene has distributed (1C), from 2012 to 2023. Addgene’s collection of CRISPR tools has grown steadily...2023, our CRISPR guide has been viewed 1,904,462 times, and the CRISPR 101 eBook has been downloaded over...globe. Our first story focuses on a technology that has changed virtually all of biology…CRISPR.  Since ...own to share. Over the years, Addgene’s repository has grown to include many CRISPR tools.  Distributing... shipped out each year.     Figure 3: Percentage of requests per year for CRISPR plasmids categorized...

  4. Important Considerations When Using AAVs

    Type
    Blog Post
    ...graduate program at the University of Manitoba. Our lab has been utilizing chemogenetics (Designer Receptors ...Although AAV2 can transduce many tissue types, it has low expression in several brain regions where other...serotypes to infect different cell types. In the CNS, it has been shown that there is variable transduction of...from the Original Location - Axonal Transport It has been demonstrated that certain serotypes can undergo... of movement, distance traveled, etc.) (4). This has also been identified with AAV2 (5), AAV6 (6) and ...required for the onset of gene expression. However, it has yet to be determined what gene expression looks like...problem in many experiments, a group of researchers has identified this feature of AAVs as a new tool for...

  5. CRISPR-Cas14: a family of small DNA-targeting enzymes enabling high-fidelity SNP genotyping

    Type
    Blog Post
    ...studied but it is only the tip of the iceberg as it has been predicted that many others are still unknown...enzymes. In contrast to other Cas enzymes, Cas14 has not been found in bacterial genomes but only in the...add-on to a diagnostic toolkit, called DETECTR, which has been created by the authors. This kit uses Cas12 ... Cas systems, Jennifer Doudna’s lab analyzed metagenomic datasets to try to determine whether simpler ...they mined a database of microbial genome and metagenome from the Department of Energy’s Joint Genome ...

  6. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...promoter of the 5’LTR has been deleted to reduce its activity. A CMV or an EF1α promoter has been inserted in...The risk of generating RCL is low but extra care has to be taken especially if you are working with proto-oncogenes...previously used to drive expression from the LTRs) has been removed, Rev (which facilitates nuclear export...expression. In contrast, in the Tet-on system, tTA has been modified (and renamed rtTA) to bind TRE only...viral binding. This method, called spinoculation, has been found very effective for T cells transduction...to infect your cells of interest.   Advantages Disadvantages Can carry large transgenes (>8 kb) Potential... cells, it was obvious that its safety was a disadvantage particularly for clinical development. In addition...

  7. Make a Splash: Notions of Scientific Impact Are Evolving

    Type
    Blog Post
    ... even society. One obvious way to do that is and has been to publish in journals with a high “impact factor...cited articles appearing in lower-prestige journals has risen steadily from 1995 to 2013. Considering citations...online access on scientific impact Partly, the story has to do with online accessibility. It is now about ...among the most cited. And, as scientific discourse has increasingly moved online, surely times have changed...articles, Anurag Acharya and colleagues report, the percentage of citations to articles in non-elite journals...

  8. Antibodies 101: Affinity Reagents

    Type
    Blog Post
    ...also reduced.  Disadvantages Every rose has its thorns and every affinity reagent has its downsides. ...created in lab in situations where a suitable antibody has been identified but a smaller or less immunogenic...using combinatorial libraries, and high specificity has been reported for a number of targets. Due to their...use an alternative affinity reagent?  The main advantage of alternate affinity reagents is their size. ...nanostructures. Diabodies readily co-crystalize, a major advantage over antibodies (Kwon, 2019).  Single-domain antibodies...

  9. General Transfection

    Type
    Protocol
    ...10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through a 0.22 µm...L-alanyl-L-glutamine (or alternative stable glutamine such as glutaGRO, Corning 25-015-CI) Heat-inactivated FBS Low serum...L-alanyl-L-glutamine (or stable alternative such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add ...55 mL of heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots...transfection to determine what ratio gives the highest percentage of GFP positive cells. Refer to the table below...

  10. Addgene is Stopping Use of X (formerly Twitter)

    Type
    Blog Post
    ...communities and welcome suggestions in the comments.  X has moved significantly away from the functionality and...conversation in the scientific community. Addgene has been considering a gradual move off X as a response...conflict with our values. For this reason, our timeline has accelerated. We significantly reduced posting on ...Bluesky: @addgene.bsky.social; or @addgene on Instagram, TikTok, Facebook, and LinkedIn. We are also exploring...

  11. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...regulatory elements. To solve this problem, the Zhang lab has previously used SaCas9 (1053 aa/3.2 kb) with AAV ... these options (2.7 kb with 24 nt guides), so it has the potential to eclipse any of the CRISPR nucleases...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...be the best option for HDR, but this possibility has not been experimentally confirmed. Researchers have...cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA rather than ...2015). PubMed PMID: 26422227.   Potential advantages of Cpf1 Type II CRISPR systems based on Cas9 ...nt upstream of the PAM site, Cpf1 cleaves in a staggered fashion, creating a 5 nucleotide 5’ overhang 18...

  12. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...restriction enzyme, you will need to verify that it has be cloned in the correct orientation - this can be... A diagnostic restriction enzyme digest takes advantage of the fact that restriction enzymes cleave DNA...

  13. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    ...studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic tools...Simplified in vivo delivery using SPH mice CRISPRa has largely been limited to in vitro settings due to ...can be delivered to the CNS by lentivirus, which has a larger packaging capacity than AAV. Such an approach... or coding regions. Typically, each gene isoform has a distinct transcriptional start site. Because CRISPRa...circumvented by the Yang Lab with the development of a Suntag-p65-HSF1 (SPH) transgenic mouse (Zhou et al., 2018...This mouse expresses a modified version of the “Suntag” Cas9 activator that recruits up to twenty copies...used their optimized vectors to highlight a key advantage of CRISPRa over traditional cDNA overexpression...

  14. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...variant has increased signal-to-noise ratio and better ability to record subthreshold voltage fluctuations...optical pooled CRISPR screening, Paul Blainey’s lab has developed a multiplexing approach to Cas9-based pooled... stabilize the 3’ end of the pegRNA. As research has just begun with PE7, it will be exciting to see future...limitations of traditional AAVs, the Becirovic Lab has developed REVeRT, for gene delivery using reconstitution...Voltron2 voltage sensors By Brian O’Neill New viral vector preps expressing the Voltron2 voltage indicator...-2 research New viral vector preps of Voltron2 voltage sensors AAV vectors for targeting microglia The...earlier Voltron indicators (a fusion of Ace2N and HaloTag, Figure 4), they determined that an A122D mutation...

  15. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...require heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating...showed that the receptor-based fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high...recent months: Our selection of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use...performing genetics studies in organoids. The Organoid EasyTag System is a straightforward workflow for doing ... Figure 1: Example schematic from the Organoid EasyTag system, showing the workflow for inducible CRISPR... Image from Sun et al. 2021. The Organoid EasyTag workflow combines thoughtfully designed plasmid...

  16. The Strength of Story Telling

    Type
    Blog Post
    ...Show that as the story has progressed, you or your lab or science or your CV has grown as well. Find your...Task, Action, Result) interview technique, which has been addressed in much better pieces than I’m able... your structure. Develop your protagonist In most cases, the protagonist will be you. Make sure to highlight...enjoy, remember, and listen to narratives — take advantage of that in your work The part of my job I most...

  17. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ...Kcleave (min-1). SpCas9 has maximum activity from 35-45 °C, while GeoCas9 has maximum activity from 50...the recent discovery of GeoCas9 by the Doudna lab has opened up the field of thermophilic bacteria to CRISPR...stability in human plasma The last trick GeoCas9 has up its sleeve is its increased longevity in human...stable when incubated in reaction buffer alone, but has limited activity when incubated in diluted human ...50-70 °C. The key advantage of GeoCas9 is that it functions at temperatures much warmer than SpCas9...

  18. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...It is fair to say that CRISPR has revolutionized molecular biology and has enabled functional studies in...enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of Dicer enzymes and these...knockout methods of the same gene. However, recent work has called this prevalent assumption into question. Specifically...multiple target sequences are concatenated. hpRNA has been extensively used in plant research and many ...constructs are available to the community. Each construct has unique features, such as inducible expression of ...tool requires careful balancing of the inherent advantages and limitations of any technique in the context...interest. Tools for RNAi Arguably the greatest advantage of RNAi is the straightforward nature of the technique—all...

  19. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...ElizabethKenkel) December 5, 2018 How social media has helped me in science It may have taken a bit of FOMO...Here are three examples of how using social media has helped me or my lab mates: For me, using social ...with social media as a scientist Using social media has enriched my professional development, and while it...helped you fit social media into your busy schedule. Has social media helped you advance your research or ...handle, but I wasn’t using them to my professional advantage. It wasn’t until I wrote a blog post about a research...start using social media to their professional advantage, I’ve selected five resources that have tips for...

  20. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    ...Service since 2009. He has  developed a proprietary, low cost cloning protocol that he has used for cloning ...least 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region... most of the troubleshooting regarding this step has to be strategy specific, there are few general parameters...ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to the strategy adopted...satellite colonies and they form when all the antibiotic has been used. Don’t worry too much, just pick the big...many softwares out there than can help you at this stage and that can be used to simulate in silico cloning...
Showing: 261 - 280 of 520 results